LOCUS       sequence-159256-         810 bp    DNA     linear   UNA 16-APR-2025
DEFINITION  Mammalian expression of Cas9 and sgRNA scaffold.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 810)
  AUTHORS   Habashy et al
  TITLE     CRISPR/Cas9-based system for loss-of-function screening
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 810)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 16, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: lentiCRISPR v2-Blast
FEATURES             Location/Qualifiers
     source          1..810
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             226..273
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             274..297
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             307..363
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             364..762
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
ORIGIN
        1 tacaacaagc accgggataa gcccatcaga gagcaggccg agaatatcat ccacctgttt
       61 accctgacca atctgggagc ccctgccgcc ttcaagtact ttgacaccac catcgaccgg
      121 aagaggtaca ccagcaccaa agaggtgctg gacgccaccc tgatccacca gagcatcacc
      181 ggcctgtacg agacacggat cgacctgtct cagctgggag gcgacaagcg acctgccgcc
      241 acaaagaagg ctggacaggc taagaagaag aaagattaca aagacgatga cgataaggga
      301 tccggcgcaa caaacttctc tctgctgaaa caagccggag atgtcgaaga gaatcctgga
      361 ccgatggcca agcctttgtc tcaagaagaa tccaccctca ttgaaagagc aacggctaca
      421 atcaacagca tccccatctc tgaagactac agcgtcgcca gcgcagctct ctctagcgac
      481 ggccgcatct tcactggtgt caatgtatat cattttactg ggggaccttg tgcagaactc
      541 gtggtgctgg gcactgctgc tgctgcggca gctggcaacc tgacttgtat cgtcgcgatc
      601 ggaaatgaga acaggggcat cttgagcccc tgcggacggt gccgacaggt gcttctcgat
      661 ctgcatcctg ggatcaaagc catagtgaag gacagtgatg gacagccgac ggcagttggg
      721 attcgtgaat tgctgccctc tggttatgtg tgggagggct aagaattcga tatcaagctt
      781 atcggtaatc aacctctgga ttacaaaatt
//
